Drought-triggered leaf transcriptional responses disclose key molecular pathways underlying leaf water use efficiency in sugarcane (Saccharum spp.).

Drought is a major constraint to sugarcane (Saccharum spp.) production and improving the water use efficiency (WUE) is a critical trait for the sustainability of this bioenergy crop. The molecular mechanism underlying WUE remains underexplored in sugarcane. Here, we investigated the drought-triggered physiological and transcriptional responses of two sugarcane cultivars contrasting for drought tolerance, 'IACSP97-7065' (sensitive) and 'IACSP94-2094' (tolerant). After 21 days without irrigation (DWI), only 'IACSP94-2094' exhibited superior WUE and instantaneous carboxylation efficiency, with the net CO2 assimilation being less impacted when compared with 'IACSP97-7065'. RNA-seq of sugarcane leaves at 21 DWI revealed a total of 1,585 differentially expressed genes (DEGs) for both genotypes, among which 'IACSP94-2094' showed 617 (38.9%) exclusive transcripts (212 up- and 405 down-regulated). Functional enrichment analyses of these unique DEGs revealed several relevant biological processes, such as photosynthesis, transcription factors, signal transduction, solute transport, and redox homeostasis. The better drought-responsiveness of 'IACSP94-2094' suggested signaling cascades that foster transcriptional regulation of genes implicated in the Calvin cycle and transport of water and carbon dioxide, which are expected to support the high WUE and carboxylation efficiency observed for this genotype under water deficit. Moreover, the robust antioxidant system of the drought-tolerant genotype might serve as a molecular shield against the drought-associated overproduction of reactive oxygen species. This study provides relevant data that may be used to develop novel strategies for sugarcane breeding programs and to understand the genetic basis of drought tolerance and WUE improvement of sugarcane.

The plastid-nucleus localized DNA-binding protein WHIRLY1 is required for acclimation of barley leaves to high light.

MAIN CONCLUSION: In accordance with a key role of WHIRLY1 in light-acclimation mechanisms, typical features of acclimation to high light, including photosynthesis and leaf morphology, are compromised in WHIRLY1 deficient plants. Acclimation to the environment requires efficient communication between chloroplasts and the nucleus. Previous studies indicated that the plastid-nucleus located WHIRLY1 protein is required for the communication between plastids and the nucleus in situations of high light exposure. To investigate the consequences of WHIRLY1 deficiency on the light acclimation of photosynthesis and leaf anatomy, transgenic barley plants with an RNAi-mediated knockdown of HvWHIRLY1 were compared to wild-type plants when growing at low and high irradiance. While wild-type plants showed the typical light acclimation responses, i.e. higher photosynthetic capacity and thicker leaves, the WHIRLY1 deficient plants were not able to respond to differences in irradiance. The results revealed a systemic role of WHIRLY1 in light acclimation by coordinating responses at the level of the chloroplast and the level of leaf morphology.

Energy conversion processes and related gene expression in a sunflower mutant with altered salicylic acid metabolism.

Salicylic acid (SA) is involved in several responses associated with plant development and defence against biotic and abiotic stress, but its role on photosynthetic regulation is still under debate. This work investigated energy conversion processes and related gene expression in the brachytic mutant of sunflower lingering hope (linho). This mutant was characterized by a higher ratio between the free SA form and its conjugate form SA O-ß-D-glucoside (SAG) compared to wild type (WT), without significant changes in the endogenous level of abscisic acid and hydrogen peroxide. The mutant showed an inhibition of photosynthesis due to a combination of both stomatal and non-stomatal limitations, although the latter seemed to play a major role. The reduced carboxylation efficiency was associated with a down-regulation of the gene expression for both the large and small subunits of Rubisco and the Rubisco activase enzyme. Moreover, linho showed an alteration of photosystem II (PSII) functionality, with reduced PSII photochemistry, increased PSII excitation pressure and decreased thermal energy dissipation of excessive light energy. These responses were associated with a lower photosynthetic pigments concentration and a reduced expression of genes encoding for light-harvesting chlorophyll a/b binding proteins (i.e. HaLhcA), chlorophyll binding subunits of PSII proteins (i.e. HaPsbS and HaPsbX), phytoene synthase enzyme and a different expression level for genes related to PSII repair cycle, such as HaPsbA and HaPsbD. The concomitant stimulation of respiratory metabolism, suggests that linho activated a coordinate modulation of chloroplast and mitochondria activities to compensate the energy imbalance and regulate energy conversion processes.

Silicon Confers Soybean Resistance to Salinity Stress Through Regulation of Reactive Oxygen and Reactive Nitrogen Species.

Salt stress is one of the major abiotic stressors that causes huge losses to the agricultural industry worldwide. Different strategies have been adopted over time to mitigate the negative impact of salt stress on plants and reclaim salt-affected lands. In the current study, we used silicon (Si) as a tool for salinity alleviation in soybean and investigated the influence of exogenous Si application on the regulation of reactive oxygen and reactive nitrogen species and other salt stress-related parameters of the treated plants. Our results revealed that the canopy temperature was much higher in sole NaCl-treated plants but declined in Si + NaCl-treated plants. Furthermore, the chlorophyll contents decreased with sole NaCl treatment, whereas Si + NaCl-treated plants showed improved chlorophyll contents. In addition, Si application normalized the photosynthetic responses, such as transpiration rate (E) and net photosynthesis rate (PN ) in salt-treated plants, and reduced the activity of ascorbate peroxidase and glutathione under salt stress. The expression levels of antioxidant-related genes GmCAT1, GmCAT2, and GmAPX1 started to decline at 12 h after addition of Si to NaCl-treated plants. Similarly, the S-nitrosothiol and nitric oxide (NO)-related genes were upregulated in the salt stress condition but reduced after Si supplementation. Si application downregulated genes associated with reactive oxygen species and reactive nitrogen species and reduced enzymatic and non-enzymatic antioxidants of the treated plants. Results of the current study conclude that Si mitigated the adverse effects of NaCl-induced stress by modulating the crosstalk between antioxidants and NO scavengers. It is suggested that Si may be used in agricultural systems for alleviating salt stress.

Photosynthetic regulation in seed heads and flag leaves of sagebrush-steppe bunchgrasses.

Native sagebrush-steppe bunchgrass populations are threatened by the spread and dominance of exotic invasive annual grasses, in part due to low, episodic seed production. In contrast, the widespread exotic bunchgrass, crested wheatgrass, readily produces viable seed cohorts. The mechanisms underlying these differences are unclear. To address this, we measured seed head specific mass (g m-2) and net photosynthetic assimilation (A net) as a function of internal [CO2] (A/Ci curves) in pre- and post-anthesis seed heads and flag leaves of crested wheatgrass and four native bunchgrasses to determine if differences in allocation and photosynthetic characteristics of seed heads was consistent with differential reproductive success. Crested wheatgrass seed heads had 2-fold greater specific mass compared to the native grasses, concurrent with greater CO2-saturated photosynthesis (A max), mesophyll carboxylation efficiency (CE), and higher intrinsic water-use efficiency (WUE i ; A net/stomatal conductance (g s)), but with similar relative stomatal limitations to photosynthesis (RSL). Post-anthesis seed head A max, CE, RSL and g s decreased in native grasses, while crested wheatgrass RSL decreased and CE increased dramatically, likely due to tighter coordination between seed head structural changes with stomatal and biochemical dynamics. Our results suggest native sagebrush-steppe bunchgrasses have greater stomatal and structural constraints to reproductive photosynthesis, while the exotic grass has evolved seed heads functionally similar to leaves. This study shows elucidating reproduction-related ecophysiological mechanisms provide understanding of plant attributes that underlie restoration success and could help guide the development of native plant materials with functional attributes needed to overcome demographic bottlenecks that limit their restoration into degraded sagebrush-steppe.

Evidence of Adaptation to Recent Changes in Atmospheric CO2 in Four Weedy Species.

Seeds of three C3 and one C4 annual weedy species were collected from agricultural fields in Beltsville, Maryland in 1966 and 2006, when atmospheric CO2 concentrations averaged about 320 and 380 mol mol-1, respectively. Plants from each collection year were grown over a range of CO2 concentrations to test for adaptation of these weedy species to recent changes in atmospheric CO2. In all three of the C3 species, the increase in CO2 concentration from 320 mol mol-1 to 380 mol mol-1 increased total dry mass at 24 days in plants from seeds collected in 2006, but not in plants from seeds collected in 1966. Shoot and seed dry mass at maturity was greater at the higher growth CO2 in plants collected in 2006 than in 1966 in two of the species. Down-regulation of photosynthetic carboxylation capacity during growth at high CO2 was less in the newer seed lots than in the older in two of the species. Overall, the results indicate that adaptation to recent changes in atmospheric CO2 has occurred in some of these weedy species.

Physiological acclimation dampens initial effects of elevated temperature and atmospheric CO2 concentration in mature boreal Norway spruce.

Physiological processes of terrestrial plants regulate the land-atmosphere exchange of carbon, water, and energy, yet few studies have explored the acclimation responses of mature boreal conifer trees to climate change. Here we explored the acclimation responses of photosynthesis, respiration, and stomatal conductance to elevated temperature and/or CO2 concentration ([CO2 ]) in a 3-year field experiment with mature boreal Norway spruce. We found that elevated [CO2 ] decreased photosynthetic carboxylation capacity (-23% at 25 °C) and increased shoot respiration (+64% at 15 °C), while warming had no significant effects. Shoot respiration, but not photosynthetic capacity, exhibited seasonal acclimation. Stomatal conductance at light saturation and a vapour pressure deficit of 1 kPa was unaffected by elevated [CO2 ] but significantly decreased (-27%) by warming, and the ratio of intercellular to ambient [CO2 ] was enhanced (+17%) by elevated [CO2 ] and decreased (-12%) by warming. Many of these responses differ from those typically observed in temperate tree species. Our results show that long-term physiological acclimation dampens the initial stimulation of plant net carbon assimilation to elevated [CO2 ], and of plant water use to warming. Models that do not account for these responses may thus overestimate the impacts of climate change on future boreal vegetation-atmosphere interactions.

Genotypically Identifying Wheat Mesophyll Conductance Regulation under Progressive Drought Stress.

Photosynthesis limitation by CO2 flow constraints from sub-stomatal cavities to carboxylation sites in chloroplasts under drought stress conditions is, at least in some plant species or crops not fully understood, yet. Leaf mesophyll conductance for CO2 (gm) may considerably affect both photosynthesis and water use efficiency (WUE) in plants under drought conditions. The aim of our study was to detect the responses of gm in leaves of four winter wheat (Triticum aestivum L.) genotypes from different origins under long-term progressive drought. Based on the measurement of gas-exchange parameters the variability of genotypic responses was analyzed at stomatal (stomata closure) and non-stomatal (diffusional and biochemical) limits of net CO2 assimilation rate (AN). In general, progressive drought caused an increasing leaf diffusion resistance against CO2 flow leading to the decrease of AN, gm and stomatal conductance (gs), respectively. Reduction of gm also led to inhibition of carboxylation efficiency (Vcmax). On the basis of achieved results a strong positive relationship between gm and gs was found out indicating a co-regulation and mutual independence of the relationship under the drought conditions. In severely stressed plants, the stomatal limitation of the CO2 assimilation rate was progressively increased, but to a less extent in comparison to gm, while a non-stomatal limitation became more dominant due to the prolonged drought. Mesophyll conductance (gm) seems to be a suitable mechanism and parameter for selection of improved diffusional properties and photosynthetic carbon assimilation in C3 plants, thus explaining their better photosynthetic performance at a whole plant level during periods of drought.

Long-term exposure of Norway spruce, Picea abies (L.) Karst., to ozone in open-top chambers: IV. Effects on the stomatal and non-stomatal limitation of photosynthesis and on the carboxylation efficiency.

Rates of photosynthesis at different concentrations of CO2 (0-1850 µmol mol-1 ) were measured using one clone of Norway spruce, Picea abies (L.) Karst. exposed to three different concentrations of ozone for four seasons (1985-8) in open-top chambers. The treatments were non-filtered air+ozone (NF+), non-filtered air (NF) and charcoal-filtered air (CF). The measurements were made on shoots of four different age classes. No significant effects were found on photosynthesis or on leaf conductance to CO2 in current year shoots. In 1- to 3-yr-old shoots, leaf conductance to CO2 and rates of net photosynthesis at both 330 µmol mol-1 CO2 (Pn 330), and saturating concentrations of CO2 (Pn max), decreased with increasing shoot age and ozone concentration, although this was only significant for Pn 330 in 3-yr-old shoots. In current year shoots the intercellular concentration of CO2 was significantly higher in NF+-treated shoots compared with CF- and NF-treated shoots, while in the 2- and 3-yr-old shoots it was significantly higher in NF- and NF +-treated shoots compared with CF-treated shoots. The carboxylation efficiency significantly decreased in 2- and 3-yr-old shoots from the NF and NF+ treatments compared with shoots from the CF treatment. The gas phase limitation of photosynthesis decreased with increasing shoot age and ozone concentration.

Carbon fixation in eucalypts in the field : Analysis of diurnal variations in photosynthetic capacity.

The rate of CO2 assimilation at light saturation and an intercellular CO2 concentration of 350 µl l-1 (photosynthetic capacity), measured in leaves of Eucalyptus pauciflora, E. behriana, E. delegatensis and Acacia melanoxylon, declined over the course of cloudless days under naturally varying environmental conditions as well as under constant optimal conditions for high CO2 uptake. Since the capacity did not recover during the light period, it was different from the "midday depression" of gas exchange. The change appeared to be caused neither by the diurnal variation of total leaf water potential, by photoinhibition of redox-reaction centres in photosystems nor by changes in the intrinsic properties of Ribulose-bisphosphate carboxylase-oxygenase. The decline was more pronounced in winter than in summer. It was related to the duration of illumination or the cumulative carbon gain. It was reversible in the following dark phase, and it did not occur on changeable days with short peaks of high light.Despite the decline in photosynthetic capacity, the initial slope of the CO2 response of net photosynthesis, as obtained at low intercellular CO2 concentrations, remained constant during the day, but declined at night when photosynthetic capacity recovered. In all cases stomatal conductance varied in parallel with photosynthetic capacity. The relevance of changes in photosynthetic capacity for the intercellular CO2 concentration is discussed.